Protein crystal structure determination with the crystallophore, a nucleating and phasing agent

Abstract

Obtaining crystals and solving the phase problem remain major hurdles encountered by bio-crystallographers in their race to obtain new high-quality structures. Both issues can be overcome by the crystallophore, Tb-Xo4, a lanthanide-based molecular complex with unique nucleating and phasing properties. This article presents examples of new crystallization conditions induced by the presence of Tb-Xo4. These new crystalline forms bypass crystal defects often encountered by crystallographers, such as low-resolution diffracting samples or crystals with twinning. Thanks to Tb-Xo4's high phasing power, the structure determination process is greatly facilitated and can be extended to serial crystallography approaches.

Krypton-derivatization highlights O2-channeling in a four-electron reducing oxidase

“…7 In our previous work, native tFprA structure was solved under aerobic condition with a crystallophore derivative. 19 In contrast, the first crystal structure of this report presents tFprA (tFprA anox ) was obtained in strict anaerobic conditions after native purification. The second structure was obtained aerobically in the presence of 125 bars of Kr and is refered to as tFprA oxo-Kr .…”

Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light Scattering

“…A terbium complex, called crystallophore N • 1 or Tb-Xo4, with phasing and nucleating properties was recently described [19,20,27]. When preparing lysozyme solutions in pure milliQ water including 10 mM Tb-Xo4 for screening experiments, we observed that the addition of the compound led to the formation of a pellet composed of single lysozyme crystals at the bottom of Eppendorf tubes after 20 to 30 days in the cold room.…”

The adhesive PitA pilus protein from the early dental plaque colonizerStreptococcus oralis: expression, purification, crystallization and X-ray diffraction analysis

“…PitA Chy crystals were soaked for 20 min in mother liquor containing 100 mM Tb-Xo4 (Polyvalan Crystallophore No. 1, Molecular Dimensions; Engilberge et al, 2017Engilberge et al, , 2019 to prepare a terbium-derivatized crystal form (PitA ChyTb ).…”